Acta Veterinaria et Zootechnica Sinica

Mapping, Tissue Distribution and Polymorphism of Porcine Cellular Retinol Binding Protein Gene 2 (RBP2) and Cellular Retinoic Acid Binding Protein Gene 2 (CRABP2)

GONG Wei-hua;TANG Zhong-lin;YANG Shu-lin;CUI Wen-tao;LI Kui

2007, 38(9): 881-887. doi:

Abstract ( 1460 )

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The retinoid (vitamin A and its derivatives) plays a critical role in vision, growth, reproduction, cell differentiation, and embryonic development. Using the INRA IMpRH panel, porcine cellular retinol binding protein gene 2 (RBP2) and cellular retinoic acid binding protein gene 2 (CRABP2) were assigned to porcine chromosome 13 and 4, respectively. The entire coding sequence (CDS) of porcine CRABP2 was amplified using RT-PCR, and deduced amino acid sequence of porcine CRABP2 was compared to human corresponding protein. The mRNA distributions of the two genes in adult Wuzhishan miniature pig tissues (lung, skeletal muscle, spleen, heart, stomach, large intestine, lymph code, small intestine, liver, brain, kidney, and fat) were examined. A total of four SNPs were identified in porcine RBP2 and CRAPB2. Using PCR-RFLP method, the RBP2C117T SNP locus was analyzed in the Laiwu, Wuzhishan, Guizhou, Bama pig breeds, and Tongcheng pig population. Significant associations were found between different genotypes of RBP2-C117T with marbling score (MS.) and muscle shear force (MSF) traits. This SNP may be used as genetic marker in pig breeding and production.

Association between Hinf Ⅰ Polymorphism of Prolactin Receptor (PRLR) Gene and Performance of Dairy Cows

ZHANG Jia-lan;ZAN Lin-sen;SHEN Guang-lei

2007, 38(9): 888-892. doi:

Abstract ( 2060 )

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The polymorphism of intron 8 in prolactin receptor (PRLR) gene was detected by PCR-RFLP. A Hinf-I RFLP was detected in intron 8 and the polymorphic locus is common in all detected lines. Association between the founded Hinf-I RFLP in intron 8 and 305 days matured equivalency (ME), fat percent (F%), protein percent (P%), the first mating days (FMD) and the first calving days (FCD) was tested by least square analysis. No significant difference between Hinf-I RFLP and F%, FMD and FCD were founded in present study, but regular effects was showed. Individuals with BB genotype had lower ME (P<0.05) and higher P% (P<0.01). The additive effects of allele A was 296.00 kg/parity, 0.08, -0.14%, 4.55 d/cow, 15.10 d/cow for ME, F%, P%, FMD and FCD respectively. The dominant effects of allele A was 259.80 kg/parity, 0.07, -0.10%, 0.95 d/cow and 4.90 d/cow respectively.

Study on Isolating, RH Mapping of Bovine FABGL Gene

MA Yun;GAO Xue;REN Hong-yan;XU Shang-zhong;ZHANG Ying-han;XIN Ya-ping;GAO Shu-xin

2007, 38(9): 893-898. doi:

Abstract ( 822 )

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The blood samples from Chinese Simmental cattle was collected to extracted genomic DNA and the DNA of the bovine FABGL gene was determined and analyzed by homology cloning approach combined with PCR in this study. Sequence analysis and bioinformatics study showed that this genomic DNA contained 1925 nucleotides and 9 exons. And its base composition shows 90% and 84% identity with the corresponding pig and human, respectively. The SUNbRH panel was employed to determined the precise location of FABGL gene. Statistical analysis revealed that the bovine FABGL gene was mapped to the bovine chromosome (BTA) 23 and was closely linked to a framework marker BM47 at a 2-point LOD score of 31.05.

Study on Genetic Diversity between Mallard and Chinese Local Duck Breeds

SU Ying;CHEN Guo-hong;LONG Rui-jun;WAN Jiang-hong;MA Long

2007, 38(9): 899-906. doi:

Abstract ( 997 )

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The genetic polymorphisms of 17 microsatellites were investigated in nine indigenous duck breeds and one species of wild duck (mallard) in China Average effective number of alleles (Ne) and average rates of homozygote of each breeds were counted. Accordingly, allele frequencies of 17 microsatellite, polyomerphism information content (PIC), mean heterozygosity (H) and genetic distances (Ds) were also calculated. Moreover, dendrograms using UPGMA and the Neighbor-joining method were produced. The nine indigenous duck breeds and one species of wild duck (mallard) population have a high average PIC（0.650 6）and H（0.689 6）. The effective number of alleles is 4.1±1.5. Ds is between 0.163 1 and 0.684 4 among nine indigenous duck breeds. It is deduced that Chinese ducks have not only the same ancestor and about five diverges: 1) Beijing duck, Jingjiang duck; 2) Zhongshan, Jianchang and Gaoyou duck; 3) Jingxi duck, Jinding duck; 4) Liancheng duck; 5) Shaoxing duck

Polymorphism Analysis on Coding and Regulation Regions of Growth Hormone Gene in Duck

XU Sheng-hai;BAO Wen-bin;CHENG Jin-hua;HUANG Jun;ZHANG Hong-xia;CHEN Guo-hong

2007, 38(9): 907-912. doi:

Abstract ( 982 )

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The single nucleotide polymorphism (SNP) of growth hormone gene was investigated in various breeds of duck, including Beijing ducks, Xihu mallards, Cherry Valley Meat ducks, Jinding ducks, Shan Partridge ducks, Jingjiang ducks, Shaoxing ducks and Jinyun Partridge ducks. Eight pairs of primers for coding and regulation regions of GH gene were designed based on the database of duck genomic sequence and the SNPs were detected by PCRSSCP method Three SNPs were found among individuals, which were 230(C→G), 244(C→A) in 5′-UTR and 3 701(C→T) in exon 4， respectively. However, the mutation at 3 701 was a silence mutation. And the 3′-UTR represented its own high conservativeness. The analysis results showed that: ⑴At locus of 5′-UTR, the frequency of B allele in Jinding ducks was significantly higher than that in other breeds. ⑵At locus of exon 4, the frequencies of genotypes in different breeds were significantly different, and the frequency of CC genotype in meattype ducks were significantly higher than that in laying-type ducks. According to the results above, we can conclude that these SNPs may have relationship with some productive traits of ducks.

Cloning and Sequence Analysis of Melanocortin Receptor-4 Gene in Goose

ZHANG Tong-yan;CHEN Hong-quan;LIANG Zhong;LI Yan-he;GAN Xiao-wei

2007, 38(9): 913-919. doi:

Abstract ( 1804 )

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Melanocortin receptor-4 (MC4R) is one of members of the family of melanocortin receptors (MC1-5R) and belongs to the family of seven transmembrane G protein coupled receptors. It can cooperate with leptin, neuropeptide Y and melanocyte-stimulating hormone to regulate body weight and feeding. A pair of primers which were designed according to the sequence of chicken MC4R gene were used to amplify the goose MC4R gene. The results showed that sequence of goose MC4R gene was 996 bp, and homologous comparison showed 95.3% and 98.5% homology at the nucleotide and amino acid levels with chicken respectively, and 75% to 79% with mammal (human, cattle and pig, etc) in nucleotide sequence. The hylogenetic tree among mammalian, birds and fish showed goose earlier differentiated from mammals. The hydrophilicity, averagy charge and surface probability of amino acid residues varied with MC4R seven transmembrane process.

β-cypermethrin Regulating the Testosterone Synthesis of Male Mouse through ERK1/2 Cascade

WANG Xian-zhong;WU Jian-yun;SUN Yan;LUO Ying;LI Guan-hua;YUAN Zun-xue;WANG Xi-ming;ZHANG Jia-hua

2007, 38(9): 920-925. doi:

Abstract ( 847 )

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To study the effects of β-cypermethrin on the testosterone synthesis. Matured kunbai mouse were fed different concerntration of β-cypermethrin for 7 days RT-PCR, Western blot and radioimmunoassay(RIA) were used to analyze the ability of testosterone synthesis.The results were as followed: with the increase of β-cypermethrin concerntration,the weight of testis, the expression of P450scc mRNA ,3β-HSD mRNA and StAR mRNA had no difference between different groups.However, the expression of P450c17 mRNA and the level of StAR protein decreased significantly and the activity of ERK1/2 increased significantly. These results inferred that β- cypermethrine could inhibit the synthesis of testosterone through increasing the activity of ERK1/2 to decrease the activity of P450c17 mRNA and level of StAR Protein

In vitro Fermentation Kinetic Analysis of the Carbohydrate Fractions in Various Sections of Corn Stalks

XUE Hong-feng;YAN Gui-long;MENG Qing-xiang

2007, 38(9): 926-933. doi:

Abstract ( 1300 )

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The aim of this experiment was to evaluate the fermentation kinetics of carbohydrate fractions (A, B1, B2 and NDS) of 7 various sections of corn stalks of “Nongda 108”, including leaf blade (LB), leaf sheath (LS), stem bark (SB), stem pith (SP), stem node (SN), whole stalk (WS) and ear husk (EH). The results showed that carbohydrate fractions were significantly different (P < 0.000 1) among various sections of corn stalks. Content of carbohydrate fractions A from high to low rank as follows: SB > SN > LB > WS > LS > EH > SP; content of carbohydrate fractions B1 as follows: EH > LS > WS > SN > LB > SP > SB; SP had the highest content of carbohydrates fraction B2, while SB had the lowest content. Gas productions of various sections of corn stalks for carbohydrates three fractions with rank from high to low values were consistent with their contents. SB and SN had the highest content of neutral detergent soluble (NDS), while SP had the lowest content, however, SN had the highest gas production, followed by EH, and SP content were the lowest. The curve subtraction technique represents a valid approach to evaluate the fermentation kinetics of carbohydrate fractions of various sections of corn stalks.

Effects of Dietary Energy Level on the Content of Intramuscular Fat and mRNA Expression for Fatty Acid Synthase and Hormonesensitive Lipase in Growingfinishing Pigs

LIU Zuo-hua;YANG Fei-yun;KONG Lu-jun;ZHOU Xiao-rong;GU Yu-hong;WANG Xiao-you

2007, 38(9): 934-941. doi:

Abstract ( 811 )

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Seventytwo Landrace × Rongchang crossbred barrows, with initial body weight of 20.7 kg, were allocated to three treatments to study the effect of energy levels of dietary on mRNA expression for the content of intramuscular fat and their association with fatty acid synthase (FAS) and hormone-sensitive lipase(HSL) at finishing stage. The digestible energy levels of dietary were 11.75, 13.05 and 14.36 MJ/kg, respectively. The pigs were slaughtered at 100 kg body weight. The results showed that: ①There were no significant effects of energy density on average daily gain(ADG) and average daily feed intake(ADFI) (P>0.05), while feed conversion ratio was improved(P<0.05)and the duration of finishing was shortened (P < 0.05) with energy density increasing. The percentage of external fat, backfat thickness and intramuscular fat were increased (P < 0.05) with increasing of energy density; ②The dietary energy density of 14.36 MJ/kg improved the mRNA expression of FAS in backfat (P < 0.05) , decreased the mRNA expression of HSL (P < 0.05) and increased the ratio of mRNA of FAS to HSL (P < 0.05) in both backfat and intramuscle; ③There were significantly positive correlations (P < 0.05) between energy levels and intramuscular fat, FAS mRNA and the ratio of mRNA of FAS to HSL, between the ratio of mRNA of FAS to HSL and intramuscular fat . However, the extremely significantly negative correlations (P < 0.01)were observed between HSL mRNA and dietary energy，significantly negative correlations (P < 0.05) were observed between HSL mRNA and intramuscular fat. This study indicated that higher energy level of dietary could increase fat deposition. Moreover, effects of dietary energy level on fat deposition, especially on intramuscular fat deposition, maybe modulate both the mRNA of FAS and HSL, not anyone of them.

Research on the Invading Process and Distribution of Duck Enteritis Virus in Artificially Infected Ducks by Indirect ImmunoFluorescent Staining Method

CHENG An-chun;HAN Xiao-ying;WANG Ming-shu;YUAN Gui-ping;XU Chao;LIAO Yong-hong

2007, 38(9): 942-946. doi:

Abstract ( 740 )

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Fifty adult ducks were inoculated with a virulence duck enteritis virus(DEV, strain CHv）, and acute cases of duck viral enteritis(DVE) were duplicated Tissue samples from heart, liver, spleen, lung, kidney, thymus, duodenum, bursa of Fabricius, brain, esophagus and the pancreas were collected after inoculation and paraffin slices of these samples were prepared. The method of indirect immuno-fluorescent staining（IFA） was applied to investigate the invading process and distribution of DEV. The results showed that DEV antigen can be detected in the spleen, thymus and bursa of Fabricius 4 hours after infection and in the liver, esophagus, duodenum, rectum and the lung 6 hours after infection. The positive rate of viral antigen in different tissues detected by IFA was 46/50 in the liver, 48/50 in the spleen, 46/50 in the lung, 0/50 in the kidney, 46/50 in the intestine, 46/50 in the bursa of Fabricius, 47/50 in the thymus, 0/50 in the pancreas, 0/50 in the brain, 44/50 in the esophagus and 0/50 in the heart. The research suggested that the spleen, bursa of Fabricius, thymus, esophagus, intestine, liver and the lung are the main target organs of DEV in acute DVE cases. After infection, DEV appears in the spleen, the thymus and bursa of Fabricius first, then spreads to the liver, alimentary canal and the lung quickly. IFA is a sensitive and specific method in detecting DEV antigen in paraffin slices and it is a good method for diagnosis and antigen location of DEV.

Phylogentic Analysis of the Cryptosporidium Based on 18S rRNA Gene and HSP70 Gene Sequences

WANG Jin-chan;JIAN Fu-chun;ZHANG Long-xian;NING Chang-shen;YAN Wen-chao;SUN Ming-fei;QIU Shu-xing;LU Qing-bin

2007, 38(9): 947-953. doi:

Abstract ( 793 )

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In order to investigate species or genotype of the Cryptosoridia isolates that derived from different hosts in Henan province, the author amplified the complete sequences of the small-subunit rRNA ( 18S rRNA) gene and the 70 ku heat shock protein (HSP70) gene. Phylogentic analyses were performed using PAUP 4.0 and TREEPUZZLE 4.1.Species and genotypes of all Cryptosporidium isolates that derived from livestock, poultry and human in Henan were determined for understanding the mechanism of Cryptosporidium molecular epidemiology. We characterized the 18S rRNA and HSP70 genes of the isolates. Results of the phylogenetic analysis revealed that the Cryptosporidia human strain is Cryptosporidium parvum mouse genotype; the Cryptosporidia deer (wapiti) strain is C. parvum deer genotype, the two Cryptosporidia swine strains both are C. suis; the two Cryptosoridia quail strains belong to different species, one is C. bailey, the other is C．meleagridis; both the other Cryptosporidia birds strains (isolated from chicken and ostrich) are C. baileyi; the Cryptospridia isolates from cattle is C. andersoni.

Construction of Immune Regulatory DNA Vaccines for Haemonchus contortus and Their Expressions in Goats

YAN Ruo-feng;ZHAO Guang-wei;XU Li-xin;SUN Yan-ming;LI Xiang-rui

2007, 38(9): 954-958. doi:

Abstract ( 831 )

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To construct immune regulatory DNA vaccines for Haemonchus contortus, three gene fragments encoding parts of antigen H11, named as H11-1, H11-2 and H11-3, were ligated with goat interleukin-2 gene one by one, and the fusions were expressed in vector pcDNA- In order to analyze the expression of the immune regulatory DNA vaccines in vivo, lambs were administered with the mixture of pcDNA/IL-2-H11-1, pcDNA/IL-2-H11-2 and pcDNA/IL-2-H11-3. Goat muscles were taken from the injected site, the transcriptions and translations of vaccines were tested by RT-PCR and Western Blot analysis, seven days post primary immunization and ten days after boosting. The results showed that immune regulatory DNA vaccines carrying H11 and IL-2 genes were expressed in goat muscle successfully.

Effects of Estrogen on Expression of ER, NGF and ChAT in Cerebellum of Ovariectomized Female Sprague-Dawley Rat

LUO Qi-hui;CHEN Zheng-li;FAN Guang-li

2007, 38(9): 959-965. doi:

Abstract ( 827 )

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The effects of estrogen on the expression of ER, NGF and ChAT in rats cerebellum were studied in this paper.The model of ovariectomized female rat was established to study the expression and distribution of estrogen receptor (ER), nerve growth factor (NGF) and choline acetyltransferase (ChAT) in the cerebellum after 17β-estradiol treatment by using technique of immunohistochemical ultrasensitive SP in Sprague.Dawley ratThe immunoreactive products were distributed in Purkinje′s cell, dentate nucleus, interstitial nucleus and fastigial nucleus of cerebellum, and the ER positive products mainly located in plasma, cytoplasmic membrane and neurite, also existed in nucleus.The general tendency of the expression of ER, NGF and ChAT positive products in cerebellar cortex and nuclei of ovariectomized rat is significantly decreased, while the intensity and quantity of the immunoreactive products ascended predominantly after 17β-estradiol treatment. The above results suggested that the estrogen upregulated the expression of NGF and ChAT. Furthermore, the similarity of their changing tendency implied that they were correlated and cooperated during the course in effect of estrogen on cerebellum. It also showed that the action of estrogen in cerebellum could via genomic and nongenomic mechanism.

Localization of Doublelabeled Immunoreactive Cells of Gonadotropin Releasing Hormone and Oxytocin in Hypothalamus of Dairy Goat during Pregnancy

CHEN Shu-lin;LIN Lei;JIANG Tian-yuan;LEI Zhi-hai;LI Wen-xian;ZENG Ming-hua;ZHANG Yan;FAN Guang-li;SUI Zhi-guo

2007, 38(9): 966-971. doi:

Abstract ( 840 )

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In the present study, doublelabeled immunohistochemistry were adopted to explore co-localization of gonadotropin releasing hormone (GnRH) and oxytocin (OT) in cells of hypothalamus of dairy goat during pregnancy. The results showed that neurons which were double positive with both antigens were localized in nuclei such as paraventricular nucleus, supraoptic nucleus, suprachiasmatic nucleus, arcuate nucleus, lateral hypothalamic area, medial mammillary nucleus and posterior mammillary nucleus.The number of these neurons was significantly changed during different period of gestation. These results provide morphological evidence for a direct modulation between GnRH and OT systems.

Effect of Monochromatic Light on Morphology of Testis in Broilers

CAO Jing;CHEN Yao-xing;WANG Zi-xu;LI Jun-ying;XIE Dian;JIA Liu-jun;Li Yan-hong

2007, 38(9): 972-976. doi:

Abstract ( 850 )

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Using LED (lightemitting diodes) as light sources, AA (Arbor Acres) broiler’s morphology of testes were compared with the monochromatic light and discuss the internal mechanism.Day-old 48 male broilers were divided into four light treatments (n=12), including red (660 nm), green (560 nm), blue (480 nm) and control white (400-760 nm) by using LEDFeed and water were provided for ad libitum consumption.Light intensity was 15 lx at the height of birds’ heads and scheduled for 23 h of light and 1 h of dark during the experiment. Serum testosterone and thyroid hormone concentration were determined by using radioimmunoassay and testis weight, testis volume, seminiferous tubule area and the percentage of germ cells were measured at 49 days of age. The results showed that testes developments were affected significantly on blue and green light.Body weight, testis weight, testis volume, seminiferous tubule area and the percentage of germ cells in blue and green light groups were significant higher than those of red and white light groups (7.22%-32.37%, P<0.05), and thyroid hormone and testosterone level in blue were higher than those of others (14.36%-26.20%).In conclusion, blue light could be more efficient to stimulate thyroid hormone and testosterone than other monochromatic lights and to affect morphology of testis in broilers.

The Influence of Fluoride on Cell Cycle and Apoptosis of Cultured Caprine Osteoblasts

ZHONG Dai-bin;FENG Li-fang;QU Wei-jie;SU Jing-liang;HAN Bo

2007, 38(9): 977-982. doi:

Abstract ( 1621 )

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In order to determine whether fluoride would induce apoptotic cell death, our research group monitored DNA fragmentation and the cellular morphology with Hoechst33342/PI double staining，DNA ladder and transmission electron microscope (TEM). The level of apoptosis was analyzed with a fluorescenceactivated cell sorter (FACS) by the Annexin-V-FITC and the propidium iodine (PI) double staining method. Cell-cycle analysis was also examined with FACS using PI staining. The experimental results were as follows:Ultrastructural features of osteoblasts in control group exhibited clear nuclear envelope,large nucleolus, even karyoplasms, abundant endoplasmic reticulum and mitochondria. When fluoride was more than 1.0×10－4 mol/L, cellular membrane disintegration, cytoplasm condensation, chromatin compaction or fragmentation, endoplasmic reticulum (ER) expansion, nucleus shrinkage or periphery were observed Fluoride at 1.0×10^－5-2.5×10^－3 mol/L induced apoptosis with DNA fragmentation. FACS cell-cycle analysis demonstrated that fluoride caused potent G0/G1 arrest. Few cells could be found either in the S phases or the G2/M, thus inhibiting the transformation from S phase into G2/M phase under high levels of fluoride. The highest apoptosis rate of earlier and later stages were induced by 1.0×10^-4 mol/L fluoride(3.33% and 2.92% respectively).In conclusion, the results revealed that fluoride can induce apoptosis in osteoblast.

Antiabortive Effect of Baicalin and Its Impact on Cytokines in Mouse

MA Ai-tuan;ZHONG Xiu-hui;MENG Li-gen;LIU Zhan-min;ZHANG Tie;ZHOU Bang-hui

2007, 38(9): 983-988. doi:

Abstract ( 884 )

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To investigate the effect of Baicalin on abortion model and its impact on cytokines at the maternal-fetal interface,KM mice were divided into 5 groups and all mice except the control were injected with Bromocriptine subcutaneously on days 5 to 7 of pregnancy to establish abortion model. Then the mice in different groups were gavaged with distilled water, 0.1, 0.5, 1 mg Baicalin respectively on days 1 to 7 of gestation while the control group treated with ethanol and distilled water. Samples were collected on day 3, 5, 7, 10 and 12 to calculate abortion rate. The content of interferon (IFN)-γ, interleukin (IL)10, progesterone in serum and uterine supernatant were measured by Enzyme-Linked Immuno-absorbent Assay (ELISA). Results showed that the abortion rate in 1 mg Baicalin group at day 12 of gestation was similar to that of control group, which was significantly different from all the abortion models. The volume of IFN-γ was significantly increased in all groups especially in 1mg Baicalin group throughout the implantation stage (day 5 of gestation). As the gestation progresses, the content of progesterone increased gradually with the significant higher level in 0.5 mg and 1 mg Baicalin groups. These results indicate that Baicalin upregulates the level of progesterone in a dosedependent manner, enhances the secretion of IFN-γ during implantation and down-regulates the IFN-γ production afterwards. These findings suggest that Baicalin can modulate cytokine network in pregnancy and benefit embryo implantation and survival.

A Strategy to Confirm the Genetic Effect of Candidate Gene in a Mendelian Segregating Population

JIANG Run-shen;LI Jun-ying;YANG Ning

2007, 38(9): 989-992. doi:

Abstract ( 807 )

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The accuracy and the reliability of the association between candidate gene variations and target traits are influenced by the genetic background of papulation, polymorphism of genes, linkage disequilibrium of QTL and the sample size. The association, therefore, should be further confirmed in other populations. However, for the gene with low allelic frequency, it is laborious and cost-consuming to develop a large experimental population to provide adequate individuals with double minor alleles. In the present study, a strategy crossing between heterozygotes was introduced to generate a Mendelian segregating population with larger sample size of the minor homozygous individuals, and accuracy of gene genetic effect evaluation was enhanced due to the closer genetic background and the consideration of family information As an example, in this study, the genetic effect of the pituitary-specific transcription factor gene (POU1F1-6) on chicken growth rate was confirmed by this strategy.The results indicated that the strategy was applicable, laborsaving, and cost-effective.

Effect of Immunosuppressant Cyclophosphamide on Performance and Incretion in Broiler

DONG Xiao-fang;WANG Shi-kui;SA Ren-na;ZHANG Qi;TONG Jian-ming

2007, 38(9): 993-998. doi:

Abstract ( 856 )

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In this study, the effect of immunosuppressant cyclophosphamide (CPM) on performance and incretion were investigated to discuss nutritionmetabolism characteristic in the condition of immunosuppression in broiler. 120 male commercial Arbor Acres broiler with similar body weight, randomly were divided into two groups（60×2）:Control group (C) and CPM groups (T). The birds in C and T groups were injected intraperitoneal with equiroluminal of 0.90% NaCl and CPM (10 mg/kg BW) at day 20, respectively. Other factors were same between two groups. Feed intake was recorded and full excreta were collected daily.Body weight of birds per pen were measured, at days 1, 3, 5, 8 and 13 after treatment, respectively, and blood of 4 randomly selected birds per group were obtained by heart puncture and the spleen, bursa Fabricii and thymus were weighted and these tissues at 1 day after treatment were restored in liquid nitrogen to examine the content of mRNA of IL-1β, IL-2 and IFN-γ in them.The level of GC, insulin, T3 and T4 in all serum collected were measured by radioimmunoassay (RIA) and proliferation of T and B cell were examined. The results were as follows: Compared to that of control groups, at day 1 after treatment, CPM decreased bursa Fabricii index, proliferation of B lymphocytes and the content of IL-1β mRNA in bursa Fabricii and the content of IFN-γ mRNA in bursa Fabricii and thymus significantly (P<0.05) CPM increased body weight and decreased feed intake and feed crude fat and crude protein apparent digestibility significantly (P<0.05). CPM decreased the content of GC and increased the level of insulin in serum significantly (P<0.05). Besides significant increase of body weight in T groups at day 3 after treatment, there were no significant differences of other parameter determined at other time (P>0.05) These results showed that CPM induced immunosuppression in which condition performance was improved and the incretion of GC was decreased while that of insulin was increased in broiler.

Construction of cDNA Expression Library of Eimeria tenella Sporozoites

LIN Qing;CAI Xue-peng;DOU Yong-xi;ZHAI Jun-jun;YAN Hong-bin;ZHANG Yan-ming;TIAN Guang-fu;JING Zhi-zhong

2007, 38(9): 999-1002. doi:

Abstract ( 1488 )

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Total RNA and subsequent mRNA were isolated from E tenella sporozoites A library of Oligo(dT)-primed cDNA with added directional EcoR I /Hind Ⅲ linkers was constructed and ligated to the EcoR I /Hind Ⅲ arms of the screen vector, the recombinant phage DNA was packaged by using PhageMaker packaging extracts, resulting in a primary cDNA library. The library was proved to be a good quality by the control tests. The cloning efficiency was evaluated and the length of the cDNA fragment was assayed by PCR. Using the amplified library as template DNA, a pair of primers were designed according to the sequence of the E. tenella 3-1E, then the gene was amplified by PCR. The results showed that the cDNA expression library of E. tenella sporozoites was constructed and the 3-1E gene was amplified successfully. The capacity of the cDNA library was 4×106 pfu/mL and the length of inserts was about 100-3 000 bp. It is helpful in the further study on screening related genes.

Construction of Pig IL-2 Eukaryotic Expression Plasmid pcDNA-IL-2 and Its Adjuvant Effect on pcDNA-PRRSV-SC2-ORF5 Gene Vaccine

Xininigen;CHENG An-chun;WANG Ming-shu;CHEN Xi-wen;DOU Wen-bo;LI Xue-mei;LIU Wu-mei;ZHANG Ping-ying;LIU Fei;CHEN Xiao-yue

2007, 38(9): 1003-1008. doi:

Abstract ( 897 )

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In this study, pig IL-2 eukaryotic expression plasmid pcDNA-IL-2 was constructed and its adjuvant effect was investigated. Four groups of pigs were immunized with sterilized water, empty vector pcDNA3.1(+), PRRSV-SC2-OFR5 gene eukaryotic expression plasmid pcDNA-PRRSV-SC2-ORF5 and a mixed solution of pcDNA-IL-2 and pcDNA-PRRSV-SC2-ORF5 respectively. Serum of pig in each group was collected at different time post inoculation, and antibody to PRRSV was measured by ELISA. Peripheral T lymphocytes were acquired from a small quantity of blood of each pig, and the activity of lymphocyte proliferation was detected by MTT methods after being cultured with RPMI1640 culture medium. Content of CD+4and CD+8 of peripheral T lymphocytes in each groups of pigs were tested by cytospectrometry. Results showed that the antibody and peripheral T lymphocytes level, content of CD+4,CD+8 cell of the pigs immunized with both pcDNA-IL-2 and pcDNA-PRRSV-SC2-ORF5 were significantly higher than that of the pigs immunized with pcDNA-PRRSV-SC2-ORF5 alone, indicate that plasmid pcDNA-IL-2 can be used as a adjuvant to enhance PRRS gene vaccine.

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